REGULATION OF COLLAGEN MATRIX MINERALIZATION

描述（由申请人提供）：本研究的长期目标是了解骨和牙齿中胶原基质矿化的调控机制。在上一个资助期内，我们部分表征了胶原交联的独特方面，以及矿化组织中胶原结合小亮氨酸蛋白聚糖（CB-SLRP）的特定组织分布和mRNA表达模式。这些和其他发现使我们假设：1。I型胶原的共价分子间交联参与调节矿化的空间方面，以及2.一种胶原结合蛋白聚糖，核心蛋白聚糖（DCN），部分调节胶原矿化的时间和质量。为了验证这些假设，我们想：1。建立合成低水平赖氨酰羟化酶-2的成骨细胞克隆，以将交联途径从矿化组织表型转换为非矿化组织表型，1b。表征胶原基质的赖氨酸羟基化和交联化学（类型、数量和分子分布），以及体外和体内克隆产生的矿化模式。2a.通过在成骨细胞系中过表达和反义表达DCN，研究DCN在体内外胶原成熟和矿化中的作用。2b.表征DCN缺陷和DCN/双糖链聚糖双缺陷小鼠骨/牙本质中的基质矿化（胶原和矿物质）。2c.评价DCN-胶原相互作用对体外胶原成熟的影响。从这项研究中获得的数据可能会提供深入了解骨和牙齿中胶原矿化的调节机制。

DESCRIPTION (provided by applicant): The long-term goal of this study is to understand the regulatory mechanisms of collagen matrix mineralization in bones and teeth. During the last grant period, we have partially characterized the unique aspects of collagen cross-linking, and specific tissue distribution as well as mRNA expression pattern of collagen-binding small leucine-rich proteoglycans (CB-SLRPs) in mineralized tissues. These and other findings have led us to hypothesize: 1. the covalent intermolecular cross-linking of type I collagen is involved in regulating the spatial aspect of mineralization, and 2. a collagen-binding proteoglycan, decorin (DCN), partially regulates the timing and quality of collagen mineralization. To test these hypotheses, we would like: 1a. To establish osteoblastic cell clones that synthesize low levels of lysyl hydroxylase-2 to switch the cross-linking pathway from mineralized to non-mineralized tissue phenotype, 1b. To characterize the lysine hydroxylation and cross-linking chemistries (type, quantity and molecular distribution) of collagen matrix, and the mineralization pattern produced by the clones in vitro and in vivo. 2a. To investigate the roles of DCN in collagen maturation and mineralization in vitro and in vivo by employing overexpression and antisense approaches in osteoblastic cell line. 2b. To characterize the matrix mineralization (collagen and mineral) in bones/dentin in DCN-deficient and DCN/biglycan-double deficient mice. 2c. To evaluate the effects of DCN-collagen interaction on collagen maturation in vitro. The data obtained from this study may provide insights into the regulatory mechanisms of collagen mineralization in bones and teeth.